Optimization of dose of methyltestosterone (MT) hormone for sex reversal in tilapia (Oreochromis niloticus L.)

This paper describes the optimization of dose of methyltestosteronei (MT) hormone for masculinization of tilapia (Oreochromis niloticus). Five treatments (i.e. T1 T2, T2, T4 and T5) with different doses such as 0, 40, 50, 60 and 65 mg of MT hormone were mixed with per kg of feed for each treatment and fed the fry four times a day up to satiation for a period of 30 days. The stocking density was maintained 10 spawn/liter of water. The growth of fry at different treatments was recorded weekly and mortality was recorded daily. At the end of hormone feeding the fry were reared in hapas fixed in ponds for another 70 days and at the 100th day the fish were sexed by the gonad squashing and aceto-carmine staining method. The analysis of growth data did not show any significant variation in length and weight of fish among the different treatments. High mortality of fry ranging 66% to 81.6% was observed in different treatments and highest mortality was observed during the first twelve days of the experiment. The sex ratio analysis showed that T2 (40 mg/kg) and T5 (65 mg/kg) produced 93.33% of sex reversed male and T3 (50 mg/kg) and T4 (60 mg/kg) produced 96.66% sex reversed male, and these ratios were significantly (p<0.05) different from 1:1 male: female sex ratio. The control, T1 (0 mg/kg) contained 43.33% male progeny. From these results it is suggested that either 50 mg/kg or 60 mg/kg of MT with a feeding period of 30 days could be considered as an optimum dose for masculinization of tilapia (O. niloticus).

Preliminary success on hormone induced captive breeding of goldspot mullet, Liza parsia (Ham.)

An attempt was made to breed goldspot mullet, Liza parsia in captivity through hormone induction. The fish started spawning 35-36 hours after a single dose of 2ml ova prim per kg body weight. Hatching of fertilized eggs completed within 42-48 hours after spawning. The mean hatching rate (%) was 71.33±12 corresponding to the fertilization rate (%) of 64±12. The larvae started its first external feeding on the third day and attained a length 2.5±0.25 mm. The salinity of both breeding and rearing cisterns was 20‰ and temperature was maintained at 22-23°C.

双链RNA对基因表达的抑制及其应用

RNAi(RNA interference)为研究未知功能基因提供了新的反向遗传学手段；并能应用于人类的基因治疗。文中就RNAi的研究进展、作用机制及其应用进行了评述，并与基因敲除及反义RNA抑制进行了比较。

外源性双链RNA对小鼠卵母细胞basonuclin基因表达的影响

dsRNA能阻抑卵母细胞中同源基因的表达,其作用相当于基因敲除。质粒表达的发夹环型dsRNA也可以有效降解basonuclin转录产物,这为研究basonuclin在卵母细胞发育早期的功能提供了新的手段。

Effects of exogenous double-stranded RNA on the basonuclin gene expression in mouse oocytes

In plants and less-advanced animal species, such as C.elegans, introduction of exogenous double-stranded RNA (dsRNA) into cells would trigger degradation of the mRNA with homologous sequence and interfere with the endogenous gene expression. It might represent an ancient anti-virus response which could prevent the mutation in the genome that was caused by virus infection or mobile DNA elements insertion. This phenomenon was named RNA interference, or RNAi. In this study, RNAi was used to investigate the function of basonuclin gene during oogenesis. Microinjection of dsRNA directed towards basonuclin into mouse germinal-vesicle-intact (GV) oocytes brought down the abundance of the cognate mRNA effectively in a time- and concentration-dependent manner. This reduction effect was sequence-specific and showed no negative effect on other non-homologous gene expression in oocytes, which indicated that dsRNA can recognize and cause the degradation of the transcriptional products of endogenous basonuclin gene in a sequence-specific manner. Immunofluorescence results showed that RNAi could reduce the concentration of basonuclin protein to some extent, but the effect was less efficient than the dsRNA targeting towards tPA and cMos which was also expressed in oocytes. This result might be due to the long half life of basonuclin protein in oocytes and the short reaction time which was posed by the limited life span of GV oocytes cultured in vitro. In summary, dsRNA could inhibit the expression of the cognate gene in oocytes at both mRNA and protein levels. The effect was similar to Knock-out technique which was based on homologous recombination. Furthermore, hairpin-style dsRNA targeting basonuclin gene could be produced by transcription from a recombinant plasmid and worked efficiently to deplete the cognate mRNA in oocytes. This finding offered a new way to study the function of basonuclin in the early stage of oogenesis by infection of primordial oocytes with the plasmid expressing hairpin-style basonuclin dsRNA.

Glucocorticoid receptor and protein/RNA synthesis-dependent mechanisms underlie the control of synaptic plasticity by stress

Learning and memory are exquisitely sensitive to behavioral stress, but the underlying mechanisms are still poorly understood. Because activity-dependent persistent changes in synaptic strength are believed to mediate memory processes in brain areas such as the hippocampus we have examined the means by which stress affects synaptic plasticity in the CA1 region of the hippocampus of anesthetized rats, Inescapable behavioral stress (placement on an elevated platform for 30 min) switched the direction of plasticity, favoring low frequency stimulation-induced decreases in synaptic transmission (long-term depression, LTD), and opposing the induction of long-term potentiation by high frequency stimulation, We have discovered that glucocorticoid receptor activation mediates these effects of stress on LTD and longterm potentiation in a protein synthesis-dependent manner because they were prevented by the glucocorticoid receptor antagonist RU 38486 and the protein synthesis inhibitor emetine. Consistent with this, the ability of exogenously applied corticosterone in non-stressed rats to mimic the effects of stress on synaptic plasticity was also blocked by these agents, The enablement of low frequency stimulation-induced LTD by both stress and exogenous corticosterone was also blocked by the transcription inhibitor actinomycin D, Thus, naturally occurring synaptic plasticity is liable to be reversed in stressful situations via glucocorticoid receptor activation and mechanisms dependent on the synthesis of new protein and RNA, This indicates that the modulation of hippocampus-mediated learning by acute inescapable stress requires glucocorticoid receptor-dependent initiation of transcription and translation.

RNA 的历史与未来

核酶的发现使得人们有理由相信生命起源于RNA ,通过试管演化实验获得的各种各样的催化性RNA 更使人们 对地球历史早期的RNA 世界有了越来越多的了解。同时,随着RNA 结构和功能上非凡的多样性的日益被揭示,RNA 在未来的临床应用研究中所具有的巨大潜力也正逐渐显现出来。

稳定的RNA 发夹结构在人类88 个mRNA 编码区中的分布

　以UNCG, GNRA , CUU G (N = A , U , C 或G; R = G或A) 为端环能够形成稳定的、保 守的发夹结构. 它们具有特殊的结构特征, 并在体内发挥着重要的生物学功能. 这些稳定的发夹 广泛分布于体内rRNA , 催化RNA 和非编码mRNA 中. 但对人类88 个编码区mRNA 二级结构的 研究当中, 却没有发现C(UUCG) G发夹. 而且, 与rRNA 不同, 这些编码区mRNA 四环序列的 分布没有明显的偏好性.

RNA A_6膨胀环的动力学特征

通过对RNA A6膨胀环在水溶液中的动力学模拟发现,在A6膨胀环中,环区构象主要以非堆积构象为主,环区具有较大的构象柔性,膨胀环区链的构象波动对已形成的RNA分子弯折的影响不大,弯折角只是在小范围内变动,提示作为全局结构,带大尺寸膨胀环的RNA分子仍然具有一定的刚性,柔性主要表现在膨胀环区域.

Murine MyaK, a member of a family of yeast YAK1-related genes, is highly expressed in hormonally modulated epithelia in the reproductive system and in the embryonic central nervous system

We have cloned a mouse homologue (designated Myak) of the yeast protein kinase YAK1. The 1210 aa open reading frame contains a putative protein kinase domain, nuclear localization sequences and PEST sequences. Myak appears to be a member of a growing family of YAK1-related genes that include Drosophila and human Minibrain as well as a recently identified rat gene ANPK that encode a steroid hormone receptor interacting protein. RNA blot analysis revealed that Myak is expressed at low levels ubiquitously but at high levels in reproductive tissues, including testis, epididymis, ovary, uterus, and mammary gland, as well as in brain and kidney. In situ hybridization analysis on selected tissues revealed that Myak is particularly abundant in the hormonally modulated epithelia of the epididymis, mammary gland, and uterus, in round spermatids in the testis, and in the corpora lutea in the ovary, Myak is also highly expressed in the aqueduct of the adult brain and in the brain and spinal cord of day 12.5 embryos, Mol. Reprod. Dev. 55:372-378, 2000. (C) 2000 Wiley-Liss, Inc.