Biology subjects

Kanda, S.

Publications and source records attributed to Kanda, S..

12 recordsLinked to original sources

bioRxiv2024

AgRP1 modulates breeding season-dependent feeding behavior in female medaka

Feeding and reproduction are known to be closely correlated with each other, and the seasonal breeders show breeding season-dependent feeding behavior. However, most model animals do not have definite breeding seasonality, and the mechanisms for such feeding behavior remain unclear. Here, we focused on female medaka (Oryzias latipes); they show breeding season-dependent feeding behavior, and their condition of breeding season can be experimentally controlled by day-length. We first demonstrated that, among previously reported feeding-related peptides (neuropeptides involved in feeding), agouti-related peptide 1 (agrp1) and neuropeptide y b (npyb) show higher brain expression under the breeding condition than under the non-breeding one. Combined with analysis of agrp1 knockout medaka, we obtained results to suggest that long day-induced sexually mature condition, especially ovarian estrogenic signals, increase the expressions of agrp1 in the brain, which results in increased food intake to promote reproduction. Our findings advance the understanding of neural mechanisms of feeding behavior for reproductive success.

neuroscience↗

bioRxiv2024

Inconspicuous breeding coloration to conceal eggs during mouthbrooding in male cardinalfish

The body coloration of animals has various functions, such as camouflage coloration for hiding their existence from other organisms and conspicuous coloration for appealing to their potential mates. Since the breeding colorations of males are widely considered to attract females, most previous studies on breeding coloration have mostly focused on conspicuous breeding coloration, which may have prevented the discovery of possible functions other than appealing. Here, based on a speculation that such breeding coloration might occur in species with weak sexual selection in males, we focused on Apogoninae whose sex role is considered to be reversed due to their characteristic parental behavior, paternal mouthbrooding. Through detailed morphological observations, we found that males exhibit non-conspicuous white structures, consisting of iridophores, in the lower jaw during the breeding season. Artificial implantation of eggs inside the mouth in both sexes showed that the white structure in the lower jaws, which specifically exists in males, drastically reduces the visibility of eggs during mouthbrooding. This suggested that this coloration may serve to conceal the conspicuous coloration of eggs during mouthbrooding. In addition, in vivo and in vitro hormone treatment experiments revealed that iridophore development in the lower jaw is induced by androgen through the Alkal-Ltk pathway. These results suggest that androgen-dependent breeding colorations in males, which have been considered to attract females, may serve the opposite function in these species, "inconspicuous coloration", to increase the fitness of their specialized behavior, mouthbrooding.

physiology↗

bioRxiv2023

Male-specific vasotocin expression in the medaka tuberal hypothalamus: androgen dependence and probable role in aggression

Terrestrial vertebrates have a population of androgen-dependent vasotocin (VT)-expressing neurons in the extended amygdala that are more abundant in males and mediate male-typical social behaviors, including aggression. Teleosts lack these neurons but instead have novel male-specific VT-expressing neurons in the tuberal hypothalamus. Here we found in medaka that vt expression in these neurons is dependent on post-pubertal gonadal androgens and that androgens can act on these neurons to directly stimulate vt transcription via the androgen receptor subtype Ara. Furthermore, administration of exogenous VT induced aggression in females and alterations in the androgen milieu led to correlated changes in the levels of tuberal hypothalamic vt expression and aggression in both sexes. However, genetic ablation of vt failed to prevent androgen-induced aggression in females. Collectively, our results demonstrate a marked androgen dependence of male-specific vt expression in the teleost tuberal hypothalamus, although its relevance to male-typical aggression needs to be further validated.

neuroscience↗

bioRxiv2023

Direct photoreception of a pituitary endocrine cell, melanotroph, induces a hormone release

In addition to canonical photoreception by the eye, many other organs express non-visual photoreceptors although their biological significance is mostly unknown. Here, we discovered a novel phenomenon in which the pituitary of medaka directly receives light, which induces hormone release. Ca2+ imaging analysis revealed that a melanotroph, a pituitary endocrine cell secreting melanocyte-stimulating hormone (MSH), robustly increases [Ca2+]i during short-wavelength light irradiation. Moreover, we identified Opn5m as the key molecule of this mechanism. The significance of this phenomenon was suggested to be involved in UV protection because knockout of opn5m significantly reduced the expression of tyrosinase, the rate-limiting enzyme for melanogenesis, in the skin. These results suggest a novel mechanism in which direct reception of short-wavelength light by pituitary endocrine cells triggers the pathway to enhance UV protection. One-Sentence SummaryAn endocrine cell of the pituitary was proven to be a photoreceptive cell that enables autonomous hormone release.

physiology↗

bioRxiv2023

Cholecystokinin is the follicle-stimulating hormone (FSH)-releasing hormone

In vertebrates, folliculogenesis and ovulation are regulated by two distinct pituitary gonadotropins: follicle-stimulating hormone (FSH) and luteinizing hormone (LH). Today, there is an intriguing consensus that a single hypothalamic neurohormone, gonadotropin-releasing hormone (GnRH), regulates the secretion of both FSH and LH, although the required timing and functions of FSH and LH are different. However, recent studies in vertebrates other than mammals indicate that the effect of GnRH on FSH is too weak to explain its regulation. Therefore, to challenge this "solo GnRH model," we aimed to identify the other gonadotropin regulator, FSH-releasing hormone (FSH-RH), in vertebrates. Here, by using the model teleost medaka, we successfully identified cholecystokinin as the FSH-RH. Our histological and in vitro analyses demonstrated that hypothalamic cholecystokinin-expressing neurons directly affect FSH cells through the cholecystokinin receptor, Cckbr1, thereby increasing the expression and release of FSH. Remarkably, the knockout of cholecystokinin ligand or cckbr1 minimized FSH expression and resulted in a complete failure of folliculogenesis. Our results challenge the longstanding consensus of the solo GnRH model in all vertebrates; instead, we propose the existence of a "dual GnRH model" group in vertebrates that utilizes both FSH-RH and LH-RH. The discovery of the FSH-RH in vertebrates opens not only a new era in neuroendocrinology but also possible applications involving vertebrate reproduction.

neuroscience↗

bioRxiv2023

Enhancer evolution as a driving force for lineage-specific paralog usage in the central nervous system.

Expression patterns of paralogous genes in the functionally homologous cells sometimes show differences across species. However, no reasonable explanation for the mechanism underlying such phenomena has been discovered. To understand this mechanism, the present study focused on the hypophysiotropic GnRH neurons in vertebrates as a model. These neurons express either gnrh1 or gnrh3 paralogs depending on species, and apparent switching of the expressed paralogs in them occurred at least four times in vertebrate evolution. First, we found redundant expressions of gnrh1 and gnrh3 in a single neuron in piranha and hypothesized that this situation may indicate an ancestral condition. We tested this hypothesis by examining the activity of piranha gnrh1/gnrh3 enhancers in zebrafish and medaka, in which the two gnrh paralogs are not co-expressed. Here, the gnrh1/gnrh3 enhancer of piranha induced reporter RFP/GFP co-expressions in a single hypophysiotropic GnRH neuron in both zebrafish and medaka. From these results, we propose that long-lasting ([~]550 My) redundancy after gnrh1/3 duplication in 1R/2R WGD may be the key to apparent switching of the paralog usage among the present-day species. Moreover, interspecies analyses of enhancers indicated that the loss of enhancers rather than changes in trans-regulatory elements drove the role-division of these paralogs.

evolutionary biology↗

bioRxiv2022

In vitro and in vivo gene introduction in the cloudy catshark (Scyliorhinus torazame), a cartilaginous fish

Cartilaginous fishes have various unique physiological features such as cartilaginous skeletons and a urea-based osmoregulation strategy for adaptation to their marine environment. Also, because they are considered a sister group of bony vertebrates, understanding their unique features is important from an evolutionary perspective. However, experimental approaches are limited in cartilaginous fishes. Particularly, genetic engineering, which can analyze gene functions as well as cellular behavior, has not been effectively utilized in cartilaginous fishes. This is partly because their reproductive strategy involves internal fertilization, which results in difficulty in microinjection into fertilized eggs at the early developmental stage. Trials of gene transfer have also been limited both in in vitro cultured cells and in vivo. Here, to identify efficient gene transfer methods in cartilaginous fishes, we examined the effects of various methods both in vitro and in vivo using the cloudy catshark, a candidate model cartilaginous fish species. In all methods, green fluorescent protein (GFP) expression was used to evaluate exogenous gene introduction. First, we established a primary cell culture containing fibroblast-like and epithelial-like cells from cloudy catshark embryos. Using these primary cultured cells, we attempted gene transfection by lipofection, polyethylenimine (PEI), adenovirus, baculovirus and electroporation. Among the methods tested, lipofection, electroporation and baculovirus infection enabled the successful introduction of exogenous genes into primary cultured cells, allowing us to study physiological mechanisms at a single-cell level in culture conditions close to those in a living cartilaginous fish. We also attempted in vivo transfection into cloudy catshark embryos by electroporation and baculovirus infection. Although baculovirus-injected groups did not show GFP fluorescence, electroporation successfully introduced GFP into various tissues including muscle cells. Furthermore, we succeeded in GFP introduction into adult testis by electroporation. The in vitro and in vivo gene introduction methods that worked in this study may identify paths for future genetic manipulation including knockout experiments and cellular linage analysis in cartilaginous fishes.

molecular biology↗

bioRxiv2022

Allogeneic testes transplanted into castrated adult medaka (Oryzias latipes) are engrafted and can function over a long period for production of donor-derived offspring by natural mating

Generally, successful testis transplantation has been considered to require immune suppression in the recipient to avoid rejection of the transplanted tissue. In the present study, we demonstrate in medaka that allogeneic adult testicular tissue will engraft in adult recipients immediately after partial castration without the use of immunosuppressive drugs. The allografted testes are retained in the recipients body for at least three months and are able to produce viable sperm that yield offspring after natural mating. Some recipients showed a high frequency (over 60%) of offspring derived from spermatozoa produced by the transplanted testicular tissue. Histological analyses showed that allografted testicular tissues included both germ cells and somatic cells that had established within an immunocompetent recipient testis. The relative simplicity of this testis transplantation approach will benefit investigations of the basic processes of reproductive immunology and will improve the technique of gonadal tissue transplantation.

zoology↗

bioRxiv2021

Balanced release of neuropeptide FF and gonadotropin-releasing hormone 3 modulates male sexual behavior

Animals properly perform sexual behaviors by using multiple sensory cues. However, neural mechanisms integrating multiple sensory cues and regulating motivation for sexual behaviors remain unclear. Here, we focused on peptidergic neurons, terminal nerve gonadotropin-releasing hormone (TN-GnRH) neurons, which receive inputs from various sensory systems and co-express neuropeptide FF (NPFF) in addition to GnRH. Our behavioral analyses using knockout medaka of GnRH (gnrh3) and/or NPFF (npff) demonstrated that some sexual behavioral repertories were delayed, not disrupted, in gnrh3-/- and npff-/- males, while the double knockout showed normal behaviors. We also found anatomical evidence to show that both neuropeptides modulate the sexual behavior-controlling brain areas. Furthermore, we demonstrated that NPFF activates neurons in the preoptic area via indirect pathway, which is considered to induce the increase in the motivation for male sexual behaviors. Considering these results, we propose a novel mechanism by which balanced release of co-existing peptides is important for the neuromodulatory function of TN-GnRH neurons in the control of behavioral motivation. Our results may go a long way toward understanding the functional significance of peptidergic neuromodulation in response to external environments.

neuroscience↗

bioRxiv2020

Gonadectomy and blood sampling procedures in small size teleost models

Sex steroids, produced by the gonads, play an essential role in the neuroendocrine control of reproduction in all vertebrates by providing feedback to the brain and pituitary. Sex steroids also play an important role in tissue plasticity by regulating cell proliferation in several tissues including the brain and the pituitary. Therefore, investigating the role of sex steroids and mechanisms by which they act is crucial to better understand both feedback mechanism and tissue plasticity. Teleost fish, which possess a higher degree of tissue plasticity and variations in reproduction strategies compared to mammals, appear to be useful models to investigate these questions. The removal of the main source of sex steroid production using gonadectomy together with blood sampling to measure steroid levels, have been well-established and fairly feasible in bigger fish and are powerful techniques to investigate the role and effects of sex steroids. However, small fish such as zebrafish and medaka, which are particularly good model organisms considering the well-developed genetic toolkit and the numerous protocols available to investigate their biology and physiology, raise challenges for applying such protocols due to their small size. Here, we demonstrate the step-by-step procedure of gonadectomy in both males and females followed by blood sampling in a small sized teleost model, the Japanese medaka (Oryzias latipes). The use of these procedures combined with the other advantages of using these small teleost models will greatly improve our understanding of feedback mechanisms in the neuroendocrine control of reproduction and tissue plasticity provided by sex steroids in vertebrates. SUMMARYThe article describes a quick protocol to gonadectomize and sample blood from small teleost fish, using medaka (Oryzias latipes) as a model, to investigate the role of sex steroids in animal physiology.

physiology↗

bioRxiv2020

Roles of the ClC chloride channel CLH-1 in food-associated salt chemotaxis behavior of C. elegans

The ability of animals to process dynamic sensory information facilitates foraging in an ever changing environment. However, molecular and neural mechanisms underlying such ability remain elusive. The ClC anion channels/transporters play a pivotal role in cellular ion homeostasis across all phlya. Here we find a ClC chloride channel is involved in salt concentration chemotaxis of C. elegans. Genetic screening identified two altered-function mutations of clh-1 that disrupt experience-dependent salt chemotaxis. Using genetically encoded fluorescent sensors, we demonstrate that CLH-1 contributes to regulation of chloride and calcium dynamics of salt-sensing neuron, ASER. The mutant CLH-1 reduced responsiveness of ASER to salt stimuli in terms of both temporal resolution and intensity, which could disrupt navigation strategies for approaching preferred salt concentration. Furthermore, other ClC genes appeared to act redundantly in salt chemotaxis. These findings provide insights into the regulatory mechanism of neuronal responsivity by ClCs that contribute to modulation of navigation behavior.

genomics↗

bioRxiv2019

Toward Health Management of Major Labour Force Generation by Using Infection Control Countermeasures for Haematobium Schistosomiasis -assumed to be related to occupational risk- in the Republic of Malawi

BackgroundIn Malawi, haematobium schistosomiasis is highly endemic. According to previous studies, countermeasures have been conducted mainly in school-aged children. In this study, we focused on the age groups, which are assumed to be major labour force generation. Haematobium schistosomiasis is supposed to be related to occupational activities in schistosome endemic countries. MethodsWe chronologically followed the transition of schistosome egg positive prevalence before and after mass drug administration of praziquantel (MDA) by using a urine filtering examination. We also analyzed the effectiveness of urine reagent strips from the cost perspective. FindingsThe egg positive prevalence was 34.3% (95%CI: 28.5-40.5) just before MDA in June 2010 and the highest prevalence was in the age of twenties. The egg positive prevalence reduced to 12.7% (95%CI: 9.2-17.3, p<0.01) eight weeks after the first MDA and the prevalence reduced to 6.9% (95%CI: 4.6-10.0, p<0.01) after the second MDA in August 2011. The egg positive prevalence after MDA in 2013 was reduced from 3.8% (95%CI: 2.1-6.9) to 0.9% (95%CI: 0.3-3.4) and p value was 0.050. Using urine reagent strips after MDA, the positive predictive value decreased, but the negative predictive value remained high. The cost of one urine reagent strip and one tablet of praziquantel were US$0.06 and US$0.125 in 2013 in Malawi. If the egg positive prevalence is 40%, screening subjects for MDA using urine reagent strips, the cost reduction can be estimated to be about 24% -showing an overall cost reduction. ConclusionThe combination of MDA and urine reagent strips could be both a practical and cost-effective countermeasure for haematobium schistosomiasis. It is key to recognize that haematobium schistosomiasis could be considered a disease that is assumed to have some concern with occupational risk in tropical agricultural countries such as Malawi. From this point of view, it is very important to protect the health of workers; the sound labour force generation is vital for economic growth and development in these countries. Author summarySchistosomiasis is widely endemic in the tropical and subtropical countries including Malawi, and it is related that more than 300 million people suffer from associated severe morbidity. The pathway of transmission is mainly contacting infested fresh water and it is inevitable to contact fresh water through their daily activities in Malawi. Then, they are routinely exposed to the risk of schistosome infection. Previously the main targets of schistosome control were school-aged children, but our research showed main population of schistosome infection was twenties that was presumed to be major labour force. Agriculture is the dominant industry in Malawi and it can be related to be at risk of schistosome infection during agricultural work. Schistosomiasis is presumed to have occupation-related risks, we consider that schistosome control will be a valuable step-up to economic development and make a social contribution in Malawi and many low-income tropical countries. FundingThe Ministry of Education, Culture, Sports, Science and Technology of Japans scientific research grant (JSPS KAKENHI Grant Number JP23406025). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

microbiology↗