In vitro differentiation of tooth buds from embryos and adult lizards (L. gravenhorsti): An ultrastructural comparison

It is well established that the capacity for teeth to differentiate “in vitro” depends upon: (a) the age of the embryonic rudiments at the time of excision and (b) the number of cells within each tissue type which are capable of differentiating into organ culture. This paper studies ultrastructural aspects of tooth buds grown in vitro from lizard embryos and compares these characteristics with those observed in dental germs grown in situ in older lizard embryos. Moreover, we report the self-differentiation in vitro dental tissues from adult lizard and compare this phenomenon with the main features of a morphogenetic field. Our results suggest that approximately in the first third of gestation in L. gravenhorsti the dental buds has already acquired the capacity for self-differentiation in vitro. The ultrastuctural observations show that there are no significant differences between odontoblasts and ameloblasts in situ and in vitro. The tooth from “adult lizards,” isolated by combined microsurgical and enzymatic procedure and cultured in semisolid-liquid medium were also able to differentiate teeth. This phenomenon implies that self-differentiation is not rigidly determined, and that in these animals the tooth tissues represents a continuous morphogenetic field throughout the animal's life. This property is intrinsic, resides in the isolated tooth tissues, and is relatively independent of external factors. In addition, these studies indicate that the chick chorio–allantoic membrane and the semisolid-liquid culture medium supply the majority of the factors required for development of these tissues.     

Diurnal variation in LH and temporal relationships between oscillations in LH and progesterone during the luteal phase in heifers

Diurnal variation in progesterone and LH during the luteal phase and the temporal relationships between oscillations of the two hormones were studied in 10 heifers by collection of blood samples at 0100, 0700, 1300, and 1900 h each day, beginning on Day 1 (Day 0 = ovulation). Concentration of LH on Days 5-9, but not on Days 10-14, was lower (P < 0.05) at 0700 h (0.25 ± 0.02 ng/mL) than at each of the other three hours (combined, 0.32 ± 0.02 ng/mL). An oscillation was defined as an uninterrupted increase and decrease in concentrations. The number of LH oscillations/heifer with the peak at 1900 h (6.1 ± 0.7) throughout the luteal phase was greater (P < 0.01) than for each of the other hours (combined, 4.0 ± 0.2). Diurnal variation in progesterone was not detected. Only statistically defined LH oscillations were used to determine the temporal association between the peak of an LH oscillation and various components of a progesterone oscillation. On Days 5-14, the frequency of the peak of an LH oscillation occurring at the same hour as the peak of a progesterone oscillation (26/48, 54%) was greater (P < 0.0001) than at the progesterone nadir (3/48, 6%). The frequency of the LH peak occurring during increasing (11/34, 32%) and decreasing (8/25, 32%) progesterone concentrations was intermediate (P < 0.05). Results indicated the following: 1) diurnal variation occurred in LH as determined by concentration and by the hour of the peak of an oscillation; and 2) LH oscillations were temporally and positively related to progesterone oscillations.