Table 1.
Oligonucleotide sequences used in cloning and expression analysis of tilapia and catfish CREBs.
Fig 1.
Clustal W multiple alignment of tilapia and catfish CREB deduced amino acid sequences.
Fig 2.
Cladogram showing phylogenetic analysis of the Nile tilapia and catfish CREBs with other vertebrate analogs.
(Accession no.: Human A NM_004379; Human B NM_134442; Mouse C NM_001037726; Xenopus NM_001086603;Mouse A NM_009952; Mouse B NM_133828; Rat A NM_ 134443; Rat B NM_031017; Chicken NM_ 204450; Zebrafish NM_ 200909; Pig NM_001099929; Cattle NM_174285; Songbird NM_001048256).
Fig 3.
Northern blot analysis of CREBs.
(A) Expression of CREBs in tilapia ovary and testis (VOF; Vitellogenic Ovarian Follicle; FIO, Full grown Immature Ovarian Follicle; MOF, Mature Ovarian follicle). (B) Representative densitometric analysis of CREB2 and CREB3 of tilapia. Bars depicting same letter are significantly different from each other (mean ±SEM of two independent experiments). (C) Northern blot analysis of CREBs catfish ovary (P, Preparatory; PS, Pre-spawning; S, Spawning). Note: only one form of CREB (~ 1.3 kb) in tilapia testis [CREB 1] while two forms of CREB (~2.85 [CREB 2] and ~2.75 kb [CREB 3]) in tilapia ovary.
Fig 4.
Tissue distribution pattern of tilapia CREBs as determined by semi-quantitative RT-PCR.
A plasmid DNA of CREB partial cDNA was used as positive control (PC) and PCR reaction without RT was used as negative control (NC). (Ma, Marker; B, Brain; A, Adrenal; H, Heart; S, Spleen; L, Lung; I, Intestine; K, Kidney; M, Muscle; T, Testis; O, Ovary; F, Ovarian follicle).
Fig 5.
Expression of tilapia CREBs during: (A) natural ovarian cycle and (B) hCG induced -oocyte maturation as determined by RT-PCR and Northern Blot respectively. A plasmid DNA with CREB partial cDNA was used as positive control (PC) and PCR reaction without RT was used as negative control (NC) (Ma, Marker). (C) Densitometric analysis of CREB2 and CREB3 (mean ±SEM of two independent experiments).
Fig 6.
(A) Expression of catfish CREB during natural ovarian cycle (A) and hCG induced oocyte maturation in vivo (B) and in vitro (C) as determined by real-time RT-PCR.
Data are mean±SEM of two independent experiments (n = 3). Bars depicting same letter are significantly different from each other and bars depicting * are statistically significant compared to 0 hours. A plasmid DNA with CREB partial cDNA was used as positive control and PCR reaction without RT was used as negative control.