Immunolocalization of Vasa, PIWI, and TDRKH proteins in male germ cells during spermatogenesis of the teleost fish Poecilia reticulata

Milani, Liliana; Cinelli, Francesco; Iannello, Mariangela; Lazzari, Maurizio; Franceschini, Valeria; Maurizii, Maria Gabriella

doi:10.1016/j.acthis.2022.151870

Cited by 3 publications

(1 citation statement)

References 85 publications

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“…The study was performed by using *AB zebrafish (Danio rerio) and guppy (Poecilia reticulata) adult males and females, respectively, obtained from a local supplier and recently analyzed for other purposes, as described in our previous published reports [34,41]. Fish were housed in an aquarium under standard photoperiod conditions (14 h light and 10 h dark) and a temperature of 28 • C. The animals have not received any medical treatment before or during the experiments.…”

Section: Dissection Of Animals and Organsmentioning

confidence: 99%

Analysis of clasp2 Transcription Pattern in Male Germ Cells during Spermatogenesis: A Comparative Study in Zebrafish (Danio rerio) and Guppy (Poecilia reticulata)

Ricci,

Lazzari,

Maurizii

et al. 2023

Animals

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Cytoplasmic linker-associated protein-2 (CLASP2) is a member of the CLIP-associating proteins (CLASPs) family involved in the structure and function of microtubules and Golgi apparatus. Several studies performed using different mammalian and non-mammalian model organisms reported that CLASP2 controls microtubule dynamics and the organization of microtubule networks. In Drosophila and mice, an important role of CLASP2 during the development of germ cell lines has been uncovered. However, no study has clearly defined its role during fish germ cell differentiation. In the present study, we used two excellent aquatic animal models among teleost fish: zebrafish (Danio rerio) and guppy (Poecilia reticulata). Using qPCR, we found that the clasp2 transcript level is significantly high in the testis of both fish. Then, by in situ hybridization, we localized the clasp2 transcript in the spermatozoa of zebrafish and the spermatozeugmata of guppy. Our data suggest a potential role for this gene in the last stage of spermiogenesis in fish.

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Section: Dissection Of Animals and Organsmentioning

confidence: 99%

Analysis of clasp2 Transcription Pattern in Male Germ Cells during Spermatogenesis: A Comparative Study in Zebrafish (Danio rerio) and Guppy (Poecilia reticulata)

Ricci,

Lazzari,

Maurizii

et al. 2023

Animals

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Immunohistochemical Analysis of Olfactory Sensory Neuron Populations in the Developing Olfactory Organ of the Guppy, Poecilia reticulata (Cyprinodontiformes, Poecilidae)

Bettini,

Lazzari,

Milani

et al. 2023

Microscopy and Microanalysis

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Olfaction is fundamental for sensing environmental chemicals and has obvious adaptive advantages. In fish, the peripheral olfactory organ is composed of lamellae in which the olfactory mucosa contains three main categories of olfactory sensory neurons (OSNs) as follows: ciliated (cOSNs), microvillous (mOSNs), and crypt cells. We studied the appearance of these different OSNs during development of Poecilia reticulata, given its growing use as animal model system. We performed immunohistochemical detection of molecular markers specific for the different OSNs, carrying out image analyses for marked-cell counting and measuring optical density. The P. reticulata olfactory organ did not show change in size during the first weeks of life. The proliferative activity increased at the onset of secondary sexual characters, remaining high until sexual maturity. Then, it decreased in both sexes, but with a recovery in females, probably in relation to their almost double body growth, compared to males. The density of both cOSNs and mOSNs remained constant throughout development, probably due to conserved functions already active in the fry, independently of the sex. The density of calretinin-positive crypt cells decreased progressively until sexual maturity, whereas the increased density of calretinin-negative crypt cell fraction, prevailing in later developmental stages, indicated their probable involvement in reproductive activities.

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Analysis of the expression of PLZF and VASA genes and their protein relationships in mouse spermatogenic stem cells during the process of differentiation into sperm

Babatabar Darzi,

Nemati,

Azizi

et al. 2023

jct

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Germ cells Spermatogonial stem cells PLZF VASAAim: Spermatogonial stem cells (SSCs) are the basis of male spermatogenesis and fertility. SSCs are distinguished by their ability to self-renew and differentiate into spermatozoa throughout the male reproductive life and pass genetic information to the next generation. Immunohistochemical analysis showed PLZF-positive cells in the basement membrane of the seminiferous tubule. It seems that the PLZF germ cell marker is specifically expressed in the spermatogonial cells of the testis. In mice with genetic deletion of the VASA gene, males show reproductive deficiency with reduced sperm production. In this study, we looked at the expression of PLZF and VASA in spermatogenic tubules and germ cells in vivo and in vitro. Material and method: Isolation of spermatogonial stem cells, Cultivation of testis on STO feeder layer, Immunohistochemistry (IHC), Immunocytochemistry (ICC) and Fluidigm reverse transcriptase-polymerase chain reaction (RT-PCR), Network analysis of protein-protein interactions (PPI), Pathway enrichment analysis and gene analysis (GO), were used to analyze the expression of PLZF and VASA in mice testis tissue. Results: In this experimental study, whereas undifferentiated spermatogonial cells sharply express PLZF, other types of germ cells located in the seminefrous tubul were negative for this marker. In other hand, the germ cells near the basal membrane of seminefrous tubul showed expression of VASA wheras the undifferentiated germ cells located on the basal membrane were negative. The ICC analysis indicated higher expression of PLZF in the isolated undifferetiated cells in compare to the differentiated germ cells. Fluidigm real-time RT-PCR result demonstrated a significant expression (P<0.05) of VASA in the spermatogonial stem cells compared to differentiated cells and also showed expression of PLZF in undifferentiated spermatogonia. In the present experiment, after the production of SSCs under the stimulation of growth factors FGF, EGF, and GDNF, immunocytochemical staining showed a clear expression of PLZF and VASA in SSCs compared to in-vivo conditions, and VASA is less expressed in these cells. The data obtained from IHC analysis showed that VASA is expressed in the center of testicular cords. The data set related to protein-protein interaction members was used in this research due to the lack of information about PLZF expression in different stages of spermatogenesis. The study of gene expression showed that several biological and functional pathways are involved in the expression of PLZF in different stages of spermatogenesis. Conclusion: These results clearly proved the role of PLZF as a specific marker for spermatogonial stem cells, and can be beneficial for advance reserach about in-vitro differentiation of SSCs to functional sperms. ، ‫شماره‬ 2 ، ‫تابستان‬ 1402 ‫صفحات‬ ، 153 ‫تا‬ 165

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Immunolocalization of Vasa, PIWI, and TDRKH proteins in male germ cells during spermatogenesis of the teleost fish Poecilia reticulata

Cited by 3 publications

References 85 publications

Analysis of clasp2 Transcription Pattern in Male Germ Cells during Spermatogenesis: A Comparative Study in Zebrafish (Danio rerio) and Guppy (Poecilia reticulata)

Analysis of clasp2 Transcription Pattern in Male Germ Cells during Spermatogenesis: A Comparative Study in Zebrafish (Danio rerio) and Guppy (Poecilia reticulata)

Immunohistochemical Analysis of Olfactory Sensory Neuron Populations in the Developing Olfactory Organ of the Guppy, Poecilia reticulata (Cyprinodontiformes, Poecilidae)

Analysis of the expression of PLZF and VASA genes and their protein relationships in mouse spermatogenic stem cells during the process of differentiation into sperm

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