Catz DS , Fischer LM , Moschella MC , Tobias ML , Kelley DB .

19949 PHS HHS , 23684 PHS HHS , R01 NS023684 NINDS NIH HHS

	FIG. 1. Northern blot analysis of total RNA isolated from pooled laryngeal muscle from female (F) or male (M) frogs. (A) Hybridization was performed for 18 hr at 42°C with a “P-labeled PstI/EcoRl fragment of F3 (f3-500). Exposure time was 2 hr. (B) Verification that each lane of the blot contains similar amounts of RNA by reprobing the membrane with a cardiac actin probe (Dworkin-Rastl et al, 1986). Exposure was overnight.
	FIG. 2. Nucleotide and derived amino acid sequences of the f3-500 MHC cDNA clone. The sequence is aligned as in the chicken embryonic MHC cDNA clone 251 (Kavinskv et aL. 1983). * indicates the termination codon. Sequence data from this article have been deposited with the EMBL/GenBank Data Libiaries underAccession No. L01495.
	FIG. 3. Expression of LM in adult larynx of X Zuouk as determined by in situ hybridization. Digoxigenin-labeled, antisense strand of f3- 500 was used as a riboprobe. (A) Male laryngeal muscle. (B and C) Female laryngeal muscle. Big arrows indicate nuclei surrounded by the digoxigenin-labeled myosin probe. Small arrows indicate unlabeled nuclei. Scale bar: A and B, 30 Grn; C, 12 um.
	FIG. 4. LM expression in different skeletal and cardiac muscles of the frogxenopus Levis. Dark-field (left) and the corresponding bright-field (right) views are shown. All sections were hybridized using 35S-labeled antisense strand except for C and D where the sense strand of f3-500 was used. A-D, dilator laryngis; E and F, geniohyoideus internus; G and H, tibialis anterior; I and J, sternoradialis; K and L, cardiac muscle. Scale bar: 100 um.
	FIG. 5. Northern analysis of laryngeal muscle RNA from developing frogs. Lanes 2,3, and 6 are from postmetamorphic stages 2,3, and 6, respectively. Hybridization conditions are as described in the legend to Fig. 1. The blots were stained with ethidium bromide to verify that each lane contained ssmilar amounts of RNA (5 pg). Blots A and B were exposed overnight; blot C is a 72-hr reexposure of the blot shown in B.
	FIG. 6. LM expression during postmetamorphic laryngeal development as determined by in situ hybridization. Antisense probe is as described in the legend to Fig. 4. No signal was ever observed with the sense strand (see also Figs. 4C and 4D). Photomicrographs were prepared using dark-field illumination. Left panels show hemisections of male larynx; right panels show hemisections of female larynx. A and B, postmetamorphic stage (PM) 0; C and D, PMl; E and F, PM2; G and H, PM3; I and J, PM5. Note that hybridization signal is confined to muscular tissue and is not seen in cartilage or connective tissue (e.g., left-hand portion of A and B). Scale bar: 100 um.
	FIG. 7. LM expression in PM2 female and male frogs treated with DHT for 3 weeks as determined by in situ hybridization. Antisense probe is as described in the legend to Fig. 4. A and B, control PM2 females and males, respectively, raised without hormone treatment for 3 weeks. C and D, DHT-treated females and males, respectively. Scale bar: 100 Um.

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