Search bioRxiv⌕ Search

Biology subjects

Ijiri, S.

Publications and source records attributed to Ijiri, S..

4 recordsLinked to original sources

Selective activation of LH-dependent transcriptional pathways determines ovulatory follicles in the hierarchical ovary of cloudy catshark

The number of offspring produced per reproductive cycle is generally explained by a luteinizing hormone receptor (LHR)-threshold model, in which only follicles expressing sufficient levels of LHR respond to the LH surge and proceed to ovulation. Using the cloudy catshark as a model, we reveal a distinct mechanism that determines the fate of ovulatory (F1) and non-ovulatory (F2) follicles independently of LHR abundance. The cloudy catshark produces only two eggs per reproductive cycle by ovulating a pair of F1 follicles from its single ovary. Both F1 and F2 follicles express comparable levels of lhr, indicating that both retain the capacity to receive the LH signal. Nevertheless, LH stimulation selectively activated ovulation-associated responses in F1 follicles. These included progesterone production via star2 upregulation, as well as transcriptional programs including RUNX transcription factors, prostaglandin signaling (ptgs2, ptger1), and matrix metalloproteinases. Moreover, direct activation of cAMP pathway, bypassing LHR activation, induced progesterone production in F1 but not F2 follicles, indicating that the critical difference in ovulatory responsiveness lies downstream of cAMP. These results demonstrate that follicular ovulatory fate is determined, at least in part, by follicle-specific transcriptional responses to cAMP, rather than by LHR abundance alone, challenging the prevailing LHR-threshold model of ovulatory regulation in vertebrates.

physiology↗

The effects of parasitism on sex allocation of a hermaphroditic acorn barnacle

Sex allocation theory predicts the adaptive allocation of resources to male versus female reproduction in simultaneous hermaphrodites in response to individual characteristics or environmental factors. Because parasites uptake resources from their hosts, their presence could affect the sex allocation of the hosts. We investigated the effects of infestation status and infestation intensity by the rhizocephalan barnacle Boschmaella japonica on reproduction, including sex allocation, of the host intertidal barnacle Chthamalus challengeri. Feeding activity was also examined as a factor related to resource intake. Both male and female reproductive investment decreased with increasing parasite infestation, and the sex allocation of large infested hosts was more male-biased than that of large uninfested hosts. Moreover, in contrast to the model prediction that male investment does not change under resource limitation, male investment decreased in infested hosts whose resources were taken by parasites. This reduction in male investment could be explained by changes in mating group size, since infested hosts have shorter penises and consequently are able to access fewer mating partners.

evolutionary biology↗

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↗

Regulation by Progestins, Corticosteroids and RU486 of Activation of Elephant Shark and Human Progesterone Receptors: An Evolutionary Perspective

We investigated progestin and corticosteroid activation of the progesterone receptor (PR) from elephant shark, a cartilaginous fish belonging to the oldest group of jawed vertebrates. Comparison with human PR provides insights into the evolution of steroid activation of human PR. At 1 nM steroid, elephant shark PR is activated by progesterone, 17-hydroxy-progesterone, 20{beta}-hydroxy-progesterone, 11-deoxycorticosterone (21-hydroxyprogesterone) and 11-deoxycortisol. Human PR, in comparison, is activated at 1 nM steroid, only by progesterone and 11-deoxycorticosterone, indicating increased progestin and corticosteroid specificity during the evolution of human PR. RU486, an important clinical antagonist of human PR, did not inhibit progesterone activation of elephant shark PR. Cys-528 in elephant shark PR corresponds to Gly-722 in human PR, which is essential for RU486 inhibition of human PR. Confirming the importance of Cys-528 in elephant shark PR, RU486 inhibited progesterone activation of the Cys528Gly mutant PR. Compared to wild-type human PR, there was an increase in activation of human Gly722Cys PR by11-deoxycortisol and a decrease in activation by corticosterone, which may have been important in selection for the mutation corresponding to human glycine-722 PR that first evolved in platypus PR, a basal mammal.

evolutionary biology↗