ING2 (inhibitor of growth protein-2) is a member of the ING-gene family and participates in diverse cellular processes involving tumor suppression, DNA repair, cell cycle regulation, and cellular senescence. As a subunit of the Sin3 histone deacetylase complex co-repressor complex, ING2 binds to H3K4me3 to regulate chromatin modification and gene expression. Additionally, ING2 recruits histone methyltransferase (HMT) activity for gene repression, which is independent of the HDAC class I or II pathway. However, the physiological function of ING2 in mouse preimplantation embryo development has not yet been characterized previously. The expression, localization and function of ING2 during preimplantation development were investigated in this study. We showed increasing expression of ING2 within the nucleus from the 4-cell embryo stage onwards; and that down-regulation of ING2 expression by endoribonuclease-prepared small interfering RNA (esiRNA) microinjection results in developmental arrest during the morula to blastocyst transition. Embryonic cells microinjected with ING2-specific esiRNA exhibited decreased blastulation rate compared to the negative control. Further investigation of the underlying mechanism indicated that down-regulation of ING2 significantly increased expression of p21, whilst decreasing expression of HDAC1. These results suggest that ING2 may play a crucial role in the process of preimplantation embryo development through chromatin regulation.
Cryopreservation is a technique used to preserve cells for long-time storage. It is widely used in agriculture to store male gametes in liquid nitrogen. The aim of this study was to determine the optimum thawing temperature and time for samples subjected to annexin V magnetic-activated cell sorting (AnMACS) as the sperm preparation technique. Pooled semen samples from three ejaculates were divided into two groups. The treatment group was subjected both to AnMACS and to being cryopreserved, whilst the control group was cryopreserved directly without MACS. Post-thaw analysis was carried out for samples thawed at either 20°C for 13 s, 37°C for 30 s, 40°C for 7 s, 60°C for 6 s or 80°C for 5 s. Sperm kinematics, viability and capacitation status were determined for samples subjected to all thawing temperatures described. Results showed that thawing at 37°C for 13 s for MACS-processed samples was a superior option compared with other thawing procedures; there was a significant difference in P < 0.05 values for curvilinear velocity (VCL μm/s) and sperm straightness (STR %) when samples were thawed at 40°C for 7 s, with fewer capacitated spermatozoa (P < 0.05) when samples were thawed at 37°C for 30 s, 40°C for 7 s or 60°C for 6 s. Hence, we can speculate that the use of AnMACS as the sperm preparation technique can somehow enhance sperm cryosurvival rate after cryopreservation, however the fertilization potential of these cells has yet to be determined.
The optimum distance and duration of ultraviolet (UV) irradiation for the complete inactivation of African catfish Clarias gariepinus egg nucleus was investigated in this study. The UV light was suspended above the unfertilized eggs at four distances (5, 10, 20 and 30 cm) and for five durations (1, 2, 3, 4 and 5 min). Then, the irradiated eggs were activated with sperm from diploid C. gariepinus and cold shocked at 5°C for 5 min just moments before cell cleavage. Ploidy analysis was performed using karyotype chromosome counting. The results obtained suggested that the further the distance, the better the hatchability rate, however prolonged duration seemed to significantly reduced hatchability. All treatments with surviving progenies at the end of the study showed evidence of successfully diploid gynogen (2n = 56) induction at different percentages. However, the optimal protocol that gave a moderately high hatchability/survival rate and completely induced gynogens was exposure of the eggs to UV irradiation at 20 cm for 1 min. It was concluded that the distance and duration of UV irradiation affects gynogenetic induction in African catfish C. gariepinus.
Breeding and larval performance of novel hybrids from reciprocal crosses of Asian catfish Pangasianodon hypophthalmus (Sauvage, 1878) and African catfish Clarias gariepinus (Burchell, 1822) were investigated in this study. Spawning was by hormonal injection of brood fish, artificial fertilization, and incubation in triplicate aquarium tanks (0.5 × 0.5 × 0.5 m3) with continuous aeration. Reciprocal crosses (♀C. gariepinus × ♂P. hypophthalmus and ♀P. hypophthalmus × ♂C. gariepinus) had lower hatchability (≤50%) than their pure siblings (≥75%). Fish from all crosses survived until the juvenile stage but survival at 35 days post hatching (dph) was higher for pure C. gariepinus sib. ♀C. gariepinus × ♂P. hypophthalmus was observed to be less resistant to degradation of water quality than the other crosses, however it had higher body weight compared with the other crosses that showed similar performance. Morphological comparison of surviving juvenile at 35 dph, showed that all ♀P. hypophthalmus × ♂C. gariepinus and 13% of the ♀C. gariepinus × ♂P. hypophthalmus exhibited the very same morphology as that of their maternal parent species, while the other portion of the ♀C. gariepinus × ♂P. hypophthalmus cross exhibited morphological traits that were intermediate between those of both parent species. This study been the first successful attempt to hybridize both species and therefore, laid the groundwork for further studies on the aquaculture potentials of the novel hybrids.
SummaryThis study investigated the breeding parameters and embryogenic development of diploid and heat shock-induced triploid eggs of Anabas testudineus (Bloch, 1792). To this effect, broodstocks of A. testudineus were induced to spawn using the Ovaprim® hormone. After fertilization, the eggs were divided into two groups and one portion heat shocked at 41°C (for 3 min), at approximately 4 min after fertilization. Results of fertilization, hatchability, as well as the sequence and timing of embryogenic development were collated from three breeding trials. Fertilization percentages were similar in both treatments (≈90%) while hatchability was higher in the diploid eggs (79.56%) than the triploid induced eggs (50.04%). Both treatments had the same sequence of embryogenetic stages; however, the timing of development was significantly delayed in the triploids (i.e. beyond the 2-cell stages) as compared with the observations in the control group (diploid eggs). Consequently, hatching time was 5 h faster in the diploid eggs [i.e. 18 hours post fertilization (hpf)] compared with the triploid induced eggs (23 hpf). The most critical stage of embryonic development in which mass mortality occurred in the different treatments was the somite stage. The status of triploid hatchlings was affirmed using erythrocyte morphology in 2-month-old fingerlings.
The study sought to investigate the chronology of events and timing of embryogenesis, as well as breeding performances of three strains of Heterobranchus longifilis from Nigeria. Fish samples were collected from Benue River in Makurdi, Niger River in Onitsha, and Rima River in Sokoto for this study. Induced spawning of the strains was carried out so that egg development could be tracked from fertilization to hatching using a simple microscope. The microphotographs obtained showed that the embryogenesis of the strains followed a similar pattern to those of other members of the family Clariidae, however with changes occurring in the specific timing of the sequences of events (i.e. interstrain and interspecies differences). When the different strains were compared, the study noted similarities (P > 0.05) in the overall breeding performance (except for fertilization rate), survival at different stages of development, timing of embryogenesis, and larvae characteristics. The outcomes of this study, therefore, provide baseline information on what genetic improvement of the species through strain crossing can be attempted in future studies.