Journal: Scientific Reports

Article Title: Identification of accessory olfactory system and medial amygdala in the zebrafish

doi: 10.1038/srep44295

Figure Lengend Snippet: ( A ) The distribution of the transcription factor Otpa (shown in green) in the preoptic region and a posterior ventral telencephalic region (the latter indicated by a question mark) shown in a sagittal view of a 5 dpf zebrafish larva (drawn after ). Major telencephalic regions treated in the Introduction are schematically indicated. ( B ) Comparison of receptor molecules and associated G proteins on olfactory sensory neurons in zebrafish and mouse. See text for more details and references. Abbreviations: ac: anterior commissure; Ce: Cerebellum; Dm: medial zone of dorsal telencephalic area (pallium); Ha: Habenula; Po: preoptic region; poc/oc: postoptic commissure/optic chiasma; OB: olfactory bulb; PT: posterior tuberculum; T: midbrain tegmentum; TH: tuberal hypothalamus; Tel: telencephalon; TeO: tectum opticum; Vd, Vp, Vs; Vv: dorsal nucleus, postcommissural nucleus, supracommissural nucleus and ventral nucleus of ventral telencephalic area (subpallium).

Article Snippet: Staining for Otpa: Rat polyclonal antibodies against the transcription factor Otpa (Immunogen: Synthetic peptide CKKPVHPGDLAPVSDA) were manufactured by Covance (USA) and used on our brain tissue at a dilution of 1:500.

Techniques: Comparison

Journal: Scientific Reports

Article Title: Identification of accessory olfactory system and medial amygdala in the zebrafish

doi: 10.1038/srep44295

Figure Lengend Snippet: Neuroanatomical analysis and identification of the intermediate nucleus of ventral telencephalon using ( A–C,D, D’) nuclear stain (DAPI) and (A’–C’, D ,D”) immunohistochemistry for Otpa. ( E ) Lateral view of adult zebrafish brain shows level of sections shown in ( A-C, A’–C’). ( F ) Dorsal view of adult zebrafish brain shows level of sections shown in ( D– D”). Abbreviations: CC: crista cerebellaris; CCe: corpus cerebelli; Ctec: commissura tecti; D: dorsal telencephalic area; Dl: lateral zone of dorsal telencephalic area; Dm: medial zone of dorsal telencephalic area; EG: eminentia granularis; OB: olfactory bulb; LI: hypothalamic lobus inferior; LL: lateral line nerves; MO: medulla oblongata; MS: medulla spinalis; oc: optic chiasma; Pit: pituitary; Po: preoptic region; SC: spinal cord; TeO: optic tectum; TH: tuberal hypothalamus; TLa: torus lateralis; V: ventral telencephalon; Vi: intermediate nucleus of ventral telencephalon; VLo/LX: vagal lobe. I: olfactory nerve; II: optic nerve; IV: trochlear nerve; VII: facial nerve; VIII: octaval nerve; X: vagal nerve.

Article Snippet: Staining for Otpa: Rat polyclonal antibodies against the transcription factor Otpa (Immunogen: Synthetic peptide CKKPVHPGDLAPVSDA) were manufactured by Covance (USA) and used on our brain tissue at a dilution of 1:500.

Techniques: Staining, Immunohistochemistry

Journal: Scientific Reports

Article Title: Identification of accessory olfactory system and medial amygdala in the zebrafish

doi: 10.1038/srep44295

Figure Lengend Snippet: Projections after a unilateral DiI injection into the olfactory bulb of an adult zebrafish shown at four telencephalic levels from anterior ( A, A’) to posterior ( E, E’), with corresponding DAPI and fluorescent photomicrographs demonstrating tracing results. (A”) is a confocal blow-up at anterior levels detailing terminal fields and retrogradely labeled cells in the posterior zone of the dorsal telencephalon (Dp) and the dorsal nucleus of the ventral telencephalon (Vd). ( C ) shows section levels of ( A ) and ( E ). Sections ( B ) and ( D ) are the immediate caudal and rostral sections, respectively, and are not separately indicated. (D”) confocal photomicrograph shows Otpa staining in the preoptic region and the intermediate nucleus of the ventral telencephalon (Vi; insert). ( F, F’) are confocal blow ups of E’ showing Di terminals and Otpa-positive cells in Vi. Stippled double arrows indicate midline. Abbreviations: acd: dorsal part of anterior commissure; Dm: medial zone of dorsal telencephalic area; Dl: lateral zone of dorsal telencephalic area; Dp: posterior zone of dorsal telencephalic area; OB: olfactory bulb; ENv: ventral entopeduncular nucleus; LI: hypothalamic lobus inferior; lot: lateral olfactory tract; mot: medial olfactory tract; PG: preglomerular complex; Pit: pituitary; Po: preoptic region; PPa: anterior parvocellular preoptic nucleus; TeO: optic tectum; TH: tuberal hypothalamus; TLa: torus lateralis; Vd: dorsal nucleus of ventral telencephalic area; SY: sulcus ypsiloniformis; Vi: intermediate nucleus of ventral telencephalon; Vp: posterior nucleus of ventral telencephalic area; Vv: ventral nucleus of ventral telencephalic area. I: olfactory nerve; II: optic nerve.

Article Snippet: Staining for Otpa: Rat polyclonal antibodies against the transcription factor Otpa (Immunogen: Synthetic peptide CKKPVHPGDLAPVSDA) were manufactured by Covance (USA) and used on our brain tissue at a dilution of 1:500.

Techniques: Injection, Labeling, Staining

Journal: Scientific Reports

Article Title: Identification of accessory olfactory system and medial amygdala in the zebrafish

doi: 10.1038/srep44295

Figure Lengend Snippet: Neuronal connections after a unilateral DiI injection into the tuberal hypothalamus in adult zebrafish shown at three levels from anterior ( A, A’) to posterior ( C, C’; note yellow arrow at injection site) with corresponding DAPI and fluorescent photomicrographs demonstrating tracing results. (B”) Confocal photomicrograph shows retrograde tracing result in the telencephalon at the level of the intermediate nucleus of the ventral telencephalon (Vi). Note also that Dm, but not Dl, has retrogradely labeled cells (see text). ( D, D’) shows confocal blow-up of B” ( D ) and corresponding Otpa stain (D’). ( E –E”) details another injection site (yellow arrow) which is shown for DAPI, Otpa and DiI in confocal photomicrographs. ( F ) shows section levels of ( A ) and ( E ). Section ( B ) is immediately caudal to ( A ), and section ( C ) is at the same level as ( E ). Abbreviations: ATN: anterior tuberal nucleus; Dm: medial zone of dorsal telencephalic area; Dl: lateral zone of dorsal telencephalic area; Dp: posterior zone of dorsal telencephalic area; OB: olfactory bulb; E: epiphysis (pineal); ENv: ventral entopeduncular nucleus; Hd: dorsal zone of periventricular hypothalamus; Hv: ventral zone of periventricular hypothalamus; lfb: lateral forebrain bundle; LH: lateral hypothalamic nucleus; LI: hypothalamic lobus inferior; lot: lateral olfactory tract; mfb: medial forebrain bundle; mot: medial olfactory tract; PG: preglomerular complex; Pit: pituitary; Po: preoptic region; PPa: anterior parvocellular preoptic nucleus; PVO: paraventricular organ; Vd: dorsal nucleus of ventral telencephalic area; SY: sulcus ypsiloniformis; TeO: optic tectum; TH: tuberal hypothalamus; TLa: torus lateralis; TPp: periventricular part of posterior tuberculum; Vi: imtermediate nucleus of ventral telencephalon; Vp: posterior nucleus of ventral telencephalic area; Vv: ventral nucleus of ventral telencephalic area. I: olfactory nerve; II: optic nerve.

Article Snippet: Staining for Otpa: Rat polyclonal antibodies against the transcription factor Otpa (Immunogen: Synthetic peptide CKKPVHPGDLAPVSDA) were manufactured by Covance (USA) and used on our brain tissue at a dilution of 1:500.

Techniques: Injection, Retrograde Tracing, Labeling, Staining

Journal: Scientific Reports

Article Title: Identification of accessory olfactory system and medial amygdala in the zebrafish

doi: 10.1038/srep44295

Figure Lengend Snippet: ( A–A ”) Confocal photomicrographs show in addition to pERK, the nuclear stain DAPI and Otpa. ( B ) Higher power details of insert show pERK and Otpa-positive cells in confocal photography in histological material of tested fish. ( C ) Larval brain in lateral view shows section level. Abbreviations: ac: anterior commissure; CeP: cerebellar plate; DT: dorsal thalamus (thalamus); E: epiphysis; EmT: eminentia thalami; GL: glomerular layer; H: hypothalamus; Ha: habenula; INL: inner nuclear layer; lfb: lateral forebrain bundle; mdG2: mediodorsal glomerulus 2; MO: medulla oblongata; N: nucleus of the medial longitudinal fascicle; OB: olfactory bulb; on: optic nerve; P: pallium; Po: preoptic region; Pr: pretectum; PTd, PTv: dorsal, ventral part of posterior tuberculum; S: subpallium; T: tegmentum; TeO: optic tectum; TeVe: tectal ventricle; Va: valvula cerebelli; VT: ventral thalamus (prethalamus).

Article Snippet: Staining for Otpa: Rat polyclonal antibodies against the transcription factor Otpa (Immunogen: Synthetic peptide CKKPVHPGDLAPVSDA) were manufactured by Covance (USA) and used on our brain tissue at a dilution of 1:500.

Techniques: Staining

Journal: Scientific Reports

Article Title: Identification of accessory olfactory system and medial amygdala in the zebrafish

doi: 10.1038/srep44295

Figure Lengend Snippet: Both, pERK + cells, as well as double-labeled cells for Otpa and pERK were counted and analyzed in imprinted and non-imprinted zebrafish larvae, stimulated with either kin or control odor. Box plots show median (Mdn), upper and lower quartile and whiskers (maximum interquartile range: 1.5). *Indicates statistical significance p: *p < 0.05. kin = kin odor stimulus; ctr = control stimulus. n imprinted kin : 10; n imprinted ctr : 8; n non-imprinted kin : 10; n non-imprinted ctr : 9. Total cell quantity of single-labeled pERK-positive cells does not differ significantly between imprinted and non-imprinting larvae, stimulated with either kin odor or control stimulus (Kruskall-Wallis test: H(2) = 3.78, p = 0.295). Furthermore, pERK-positive and Otpa-positive double-labelled cell quantity was analyzed. A significant difference in cell number was found (H(2) = 8.579, p = 0.035) between treatments. A Mann-Whitney U test was performed to determine significant differences between two treatments. No significant differences could be detected after performing the Bonferroni correction (U = 28.5, p = 0.044, Mdn non-impr kin = 0.5, Mdn impr kin = 0; U = 22.5, p = 0.018, Mdn non-impr kin = 0.5, Mdn non-impr ctr = 0).

Article Snippet: Staining for Otpa: Rat polyclonal antibodies against the transcription factor Otpa (Immunogen: Synthetic peptide CKKPVHPGDLAPVSDA) were manufactured by Covance (USA) and used on our brain tissue at a dilution of 1:500.

Techniques: Labeling, Control, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Identification of accessory olfactory system and medial amygdala in the zebrafish

doi: 10.1038/srep44295

Figure Lengend Snippet: ( A ) Neuronal activity quantified with pERK at three successive synaptic levels from peripheral sensory olfactory sensory neurons to central nervous targets (mdG2, which is immunohistochemically identified with S100 antibody because the projections of S100 immunopositive crypt cells terminate there; Vi, which is immunohistochemically identified with Otpa antibody for many of its cell bodies) after kin odor stimulation of imprinted and non-imprinted larvae. The counted pERK activated cells were located around the mdG2 and within Vi. Red tickmarks indicate significant changes in activated cell numbers seen at each level (see text for details). Higher order (i.e. secondary) olfactory projections of mediodorsal bulb area are indicated with solid black lines (targets shared with projections of entire olfactory bulb) and dashed black lines (targets specifically attributed to mediodorsal bulbar area; see literature below and text for more details). Crypt cells are widely distributed over the entire olfactory epithelium. ( B ) Projections of adult zebrafish mediodorsal olfactory bulb area (incl. mdG2) as shown in the present paper using the lipophilic tracing substance DiI (red lines). Tracer injections into tuberal hypothalamus (TH) demonstrate also a teleostean accessory olfactory pathway via Vp/Vi. Arrowheads indicate where a projection terminates. Olfactory bulb projections shown as dashed lines to telencephalic targets are selective for mediodorsal olfactory bulb (present paper and additional data from ). Abbreviations: Cr: crypt cells; Dp: posterior zone of dorsal telencephalon; Ha: habenula; Had: dorsal part of Ha; Hav: ventral part of Ha; OB: olfactory bulb; OE: olfactory epithelium; oc: optic chiasma; on: olfactory nerve; Po: preoptic region; Tel: telencephalon; TeO: optic tectum; TH: tuberal hypothalamus; Vd, Vp, Vs, Vv: dorsal, postcommissural, supracommissural, ventral nucleus of ventral telencephalon.

Article Snippet: Staining for Otpa: Rat polyclonal antibodies against the transcription factor Otpa (Immunogen: Synthetic peptide CKKPVHPGDLAPVSDA) were manufactured by Covance (USA) and used on our brain tissue at a dilution of 1:500.

Techniques: Activity Assay

Journal: Oncology Letters

Article Title: Pathological diagnosis and treatment outcome of gastric metastases from small cell lung cancer: A case report

doi: 10.3892/ol.2019.10484

Figure Lengend Snippet: Histopathological view of gastroscopy biopsy specimen. The samples were collected on May 26, 2016. Typical carcinoma (H&E) and immunohistochemistry suggesting high expression of TTF-1, Syn and Ki67 (magnification, ×200). TTF-1, thyroid transcription factor-1; Syn, synaptophysin.

Article Snippet: The slides were incubated with primary antibodies against thyroid transcription factor-1 (TTF-1; cat. no. MAB-0677; 1:200; AXIM ® Biotechnologies, Inc.), synaptophysin (Syn; cat. no. kit-0022; 1:200; AXIM ® Biotechnologies, Inc.), marker of proliferation Ki-67 (cat. no. ZM-0166; 1:200; OriGene Technologies, Inc.), neural cell adhesion molecule 1 (NCAM1; cat. no. kit-0028; 1:200; AXIM ® Biotechnologies, Inc.), chromogranin A (CgA; cat. no. MAB-0707; 1:200; AXIM ® Biotechnologies, Inc.), cytokeratin 7 (CK7; cat. no. ZM-0069; 1:200; OriGene Technologies, Inc.), CK20 (cat. no. kit-0025; 1:200; AXIM ® Biotechnologies, Inc.) and caudal type homeobox 2 (CDX-2; cat. no. RMA-0631; 1:200; AXIM ® Biotechnologies, Inc.) overnight at 4°C.

Techniques: Immunohistochemistry, Expressing

Journal: Oncology Letters

Article Title: Pathological diagnosis and treatment outcome of gastric metastases from small cell lung cancer: A case report

doi: 10.3892/ol.2019.10484

Figure Lengend Snippet: Histopathological view of cervical lymph node biopsy specimen. The samples were collected on May 30, 2016. Typical carcinoma (H&E) and immunohistochemistry suggesting high expression of TTF-1, Syn and Ki67 (magnification, ×200). TTF-1, thyroid transcription factor-1; Syn, synaptophysin.

Article Snippet: The slides were incubated with primary antibodies against thyroid transcription factor-1 (TTF-1; cat. no. MAB-0677; 1:200; AXIM ® Biotechnologies, Inc.), synaptophysin (Syn; cat. no. kit-0022; 1:200; AXIM ® Biotechnologies, Inc.), marker of proliferation Ki-67 (cat. no. ZM-0166; 1:200; OriGene Technologies, Inc.), neural cell adhesion molecule 1 (NCAM1; cat. no. kit-0028; 1:200; AXIM ® Biotechnologies, Inc.), chromogranin A (CgA; cat. no. MAB-0707; 1:200; AXIM ® Biotechnologies, Inc.), cytokeratin 7 (CK7; cat. no. ZM-0069; 1:200; OriGene Technologies, Inc.), CK20 (cat. no. kit-0025; 1:200; AXIM ® Biotechnologies, Inc.) and caudal type homeobox 2 (CDX-2; cat. no. RMA-0631; 1:200; AXIM ® Biotechnologies, Inc.) overnight at 4°C.

Techniques: Immunohistochemistry, Expressing

Journal: Oncology Letters

Article Title: Upregulation of ECT2 is associated with transcriptional program of cancer stem cells and predicts poor clinical outcome in gastric cancer

doi: 10.3892/ol.2020.11915

Figure Lengend Snippet: Gastric tumors with higher ECT2 levels possess transcriptional traits of cancer stem cells. (A) Gene set enrichment analysis enrichment score curve demonstrated higher ECT2 expression in gastric cancer samples with highly enriched breast-cancer-progenitor-related genes. (B) BUB1 and E2F7 are strongly enriched in ECT2High gastric cancer samples. ECT2, epithelial cell transforming 2; E2F7, E2F transcription factor 7; NES, normalized enrichment score; FDR, false discovery rate.

Article Snippet: Following a brief proteolytic digestion with 0.1% trypsin at 37°C for 10 min and peroxidase blocking with 3% hydrogen peroxide solution at room temperature for 15 min, the sections were incubated with primary antibodies against: ECT2 (1:400; cat. no. 07-1364; Sigma-Aldrich, Merck KGaA), BUB1 (1:200; cat. no. DF6698; Affinity Biosciences) and E2F transcription factor 7 (E2F7; 1:200; cat. no. DF2444; Affinity Biosciences) overnight at 4°C.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Upregulation of ECT2 is associated with transcriptional program of cancer stem cells and predicts poor clinical outcome in gastric cancer

doi: 10.3892/ol.2020.11915

Figure Lengend Snippet: ECT2 expression correlates with BUB1 and E2F7 mRNA expression. A significant correlation was identified between (A) ECT2 and BUB1 mRNA expression levels and (B) ECT2 and E2F7 mRNA expression levels in gastric cancer tissues, respectively. ECT2, epithelial cell transforming 2; E2F7, E2F transcription factor 7.

Article Snippet: Following a brief proteolytic digestion with 0.1% trypsin at 37°C for 10 min and peroxidase blocking with 3% hydrogen peroxide solution at room temperature for 15 min, the sections were incubated with primary antibodies against: ECT2 (1:400; cat. no. 07-1364; Sigma-Aldrich, Merck KGaA), BUB1 (1:200; cat. no. DF6698; Affinity Biosciences) and E2F transcription factor 7 (E2F7; 1:200; cat. no. DF2444; Affinity Biosciences) overnight at 4°C.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Upregulation of ECT2 is associated with transcriptional program of cancer stem cells and predicts poor clinical outcome in gastric cancer

doi: 10.3892/ol.2020.11915

Figure Lengend Snippet: BUB1 and E2F7 protein expression in human gastric cancer tissues. (A) Representative images depicting immunohistochemical staining of BUB1 (left panel) and E2F7 (right panel) in normal tissues, and well differentiated, moderately differentiated and poorly differentiated gastric cancer tissues. Magnification, ×200. IRS of (B) BUB1 and (C) E2F7 in gastric cancer and adjacent normal tissues. Data are presented as the mean ± standard error of the mean. **P<0.01. E2F7, E2F transcription factor 7; IRS, immunoreactivity score.

Article Snippet: Following a brief proteolytic digestion with 0.1% trypsin at 37°C for 10 min and peroxidase blocking with 3% hydrogen peroxide solution at room temperature for 15 min, the sections were incubated with primary antibodies against: ECT2 (1:400; cat. no. 07-1364; Sigma-Aldrich, Merck KGaA), BUB1 (1:200; cat. no. DF6698; Affinity Biosciences) and E2F transcription factor 7 (E2F7; 1:200; cat. no. DF2444; Affinity Biosciences) overnight at 4°C.

Techniques: Expressing, Immunohistochemistry, Staining