Associations of Maternal Prenatal Drug Abuse With Measures of Newborn Brain Structure, Tissue Organization, and Metabolite Concentrations.

Importance: Increasing rates of illicit drug use during pregnancy may be associated with risk for long-term health problems in prenatally exposed children. Objective: To identify the associations of prenatal exposure to illicit drugs with organization of the newborn brain. Design, Setting, and Participants: For this cohort study, a volunteer sample of 210 illicit drug-using and nonusing mothers and their newborns was enrolled from prenatal clinics and drug abuse treatment programs in New York, New York. Enrollment, scanning, and long-term follow-up occurred from September 2004 through February 2012, and image processing and statistical analyses continued through fall 2018. In addition to 26 participants with incomplete data, a total of 64 mothers were lost to follow-up during pregnancy, and 13 newborns were lost to follow-up at birth because of perinatal complications. Exposures: Newborns were assigned to 1 of 4 primary exposure groups based on the history of most frequent maternal drug use: marijuana, cocaine, methadone maintenance, and/or heroin. Unexposed newborns were controls. Main Outcomes and Measures: Unsedated magnetic resonance imaging (MRI) of newborn brains was performed shortly after birth. Infant neurodevelopmental outcomes were assessed at age 12 months. MRI modalities included anatomical imaging, diffusion tensor imaging, T2 relaxometry, and magnetic resonance spectroscopic imaging. Infant neurodevelopmental outcomes included Bayley scales of infant development-III and Vineland Adaptive Behavior Scales. Statistical analyses were performed with results represented on the brain images. Results: Of 118 mothers, 42 (35%) were in the control group (mean [SD] age, 25.9 [6.1] years), 29 (25%) were in the cocaine group (mean [SD] age, 29.0 [6.1] years), 29 (25%) were in the marijuana group (mean [SD] age, 24.3 [5.5] years), and 18 (15%) were in the methadone and/or heroin group (mean [SD] age, 30.9 [5.7] years). Not all newborns could be scanned successfully; therefore, usable MRIs were acquired for 118 newborns from predominantly minority groups and with economically disadvantaged mothers. Anatomic abnormalities were detected in similar locations across all 3 drug exposures and included smaller volumes in the dorsal, medial, and ventral surfaces of the frontal lobe and dose-related increases in volumes in the lateral temporal lobe, dorsal parietal lobe, and superior frontal gyrus. Dose-related increases in diffusion tensor measures of tissue organization, decreases in T2 relaxometry times, and increases in spectroscopy metabolite concentrations were similar across exposures. These associations of exposures with brain measures were similar to the associations of newborn age with brain measures. The anatomic and diffusion tensor imaging measures suppressively mediated the associations of prenatal exposure with poorer 12-month infant outcomes. Conclusions and Relevance: The findings suggest that prenatal drug exposure is associated with measures of newborn brain tissue in patterns that may indicate that exposures accelerated normal fetal brain maturation, which in turn mediated the associations with poorer 12-month infant outcomes.

Fluorine-tagged 5-hydroxytryptophan to investigate amino Acid metabolism in vivo.

Auxin a plant growth hormone, has a metabolic pathway that includes molecules and enzymes like those in animal brains. In this study, tomato plant seedlings (Lycopersicon esculenta) were used to investigate the fate of fluorine-tagged 5-hydroxytryptophan (PF-5-HTP) being developed for fluorine spectroscopy and imaging. Seedlings were treated with high or low concentrations of 5-HTP or PF-5-HTP and compared with controls. Metabolites of the PF-5-HTP were quantified using a custom immunoassay for the tag. Serotonin (5-HT) levels were measured with spectrofluorometry and thin-layer chromatography. Plants in treatment conditions had serotonin levels five to six times higher than controls. PF-5-HTP served as a precursor for serotonin in a biosynthetic pathway in this plant model, providing evidence for the bioavailability of the novel molecule. The increase in serotonin in plants grown in media culture supplemented with 5-HTP or PF-5-HTP might have useful applications in pharmacology.

Exploring new compounds for functional imaging using a crayfish (Procambarus clarkia) aggression paradigm.

Covalently bonding multiple fluorine atom tags to the precursors of monoamines could provide compounds for functional imaging. Theoretically, the fluorine atoms can produce detectible signal if concentrated in vesicles inside neurons. Prior to committing more costly resources to the project, evidence was sought for uptake of the molecules into neurons in living organisms. Two 19F tag configurations of seven or nine atoms were investigated. Crayfish aggression provided a paradigm for obtaining preliminary data on the scarce new molecules. After establishing that 5-hydroxytryptophan (5-HTP) elicited serotonin-like effects, the fluorine tagged versions (PF-5-HTP) were investigated; then, the elevated aggression produced by these precursors to serotonin was blocked by coadministering fluoxetine. Treatment order effects and interrater reliability of the behavioral inventory were evaluated. Preliminary evidence that these imaging compounds are taken up into neurons obtained by studying crayfish behavior later found support using more sophisticated neuroscience techniques.

The fate of perfluoro-tagged metabolites of L-DOPA in mice brains.

A novel compound for use in magnetic resonance (MR) imaging was created by covalently bonding multiple 19F atom tags to L-DOPA. Tagging L-DOPA permits bypassing the rate-limiting factor in the biosynthesis of dopamine (DA), the conversion of tyrosine into L-DOPA. The next step in the biosynthetic pathway, the removal of the carboxyl group on the molecule by the enzyme L-aromatic acid decarboxylase (AADC), happens rapidly after L-DOPA is taken up into neurons. In order to be useful as a tool in MR imaging, the novel compound and/ or its perfluoro-tagged metabolites must accumulate in vesicles in dopaminergic neurons. We administered L-DOPA with a nine 19F atom tag (250 mg/kg) to mice pups, waited 1.5 or 3 hr, and used high pressure liquid chromatography (HPLC) to examine neural tissue samples for tagged L-DOPA and tagged DA. The isomer of L-DOPA with the tag bonded at the 5 position yielded the highest conversion to tagged DA at 1.5 hr after an i.p. injection. This study provides the first direct evidence that L-DOPA, tagged with nine fluorine atoms, is taken up into mammalian brain dopaminergic neurons where it is converted to perfluro-tagged DA. The use of these tagged compounds may make it feasible to investigate the uptake and conversion of important neurotransmitter in vivo with fluorine imaging.

Acute no-effect dose for in ova exposure to C3F7 tagged 5-hydroxytryptophan, a novel probe for investigating neural development.

PURPOSE: To create a probe for investigating neural development in chicks using magnetic resonance, a C(3)F(7) tag was bonded to the indole ring of 5-hydroxytryptophan (5-HTP). Immunoassays indicate that a low dose (5 microg/egg) administered in ova results in 0.5-1.0 microg accumulating in a 250 microL midbrain. PROCEDURES: Given the potential for developmental neurotoxicity of a compound that replaces an endogenous neurotransmitter, we investigated the effects of acute exposure during development. RESULTS: No qualitative differences were observed between treatment groups. Differences in mortality rates between treatment groups were not statistically significant [X(2)(obs)=2.30, X(2)(crit) (df=2)=5.99, P>.05]. CONCLUSION: The no-effect dose for in ova administration of C(3)F(7) tagged5-HTP is 5 microg/egg on Day 17.

T-maze performance after developmental exposure to 19F tagged 5-HTP in chicks.

Chicks were used as a model to investigate behavioral effects of administering a new compound intended for use with magnetic resonance. The compound has multiple 19F atom tags covalently bonded to the indole ring of 5-hydroxytryptophan (PF-5HTP), the immediate precursor to the neurotransmitter serotonin. On incubation Day 17, 5 microg of PF-5-HTP, an equivalent amount of 5-HTP, or just 200 microL of the weak phosphate buffered saline (PBS) vehicle was injected into the airsac of each egg. Three days after hatching, chicks were isolated at the top of a simple T-Maze which, when traversed correctly, enabled them to return to their brood mates. A second trial in the T-Maze was conducted about three hours later. The brief period of isolation at the start of a trial causes social distress in chicks who are reinforced by returning to the brood. The task was selected as being sensitive to functioning of the serotonin pathways whose development might be altered by administering the compound during brain development. Repeated-measures analysis of variance yielded a statistically significant main effect for trial within groups, but no significant difference between injection groups. Administering a low dose of the fluorine tagged compound during development did not impair performance on this T-maze task.

Tissue immunoassay for 19F-tagged 5-hydroxytryptophan.

A new tool for magnetic resonance, L-6-heptafluorobutyryl-5-hydroxytryptophan, was synthesized and investigated using an antibody to perfluoroalkyl moieties developed previously. To be useful as an imaging agent, the compound must cross the blood brain barrier and then be concentrated in vesicles in serotonergic neurons in order to accumulate in sufficient quantity for in vivo detection to be possible. The novel imaging compound was administered in ova to domestic chicks (Gallus domestics) to investigate the bioavailability and uptake dynamics of the compound in this model organism. Typical immunoassay methods were ineffective, so a new technique was developed which binds amines and amino acids to the walls of acid-functionalized cuvettes. The first study established the presence of higher quantities of the tags in neural and liver tissue than in heart tissue. A second study investigated regional differences, with the midbrain containing more tagged compounds than the frontal lobe sample, and the frontal lobe sample containing more than the occipital or cerebellum samples. These studies demonstrate that the compound follows the pathway of endogenous serotonin. A third study investigated uptake dynamics of the novel compound. Maximum concentration of the tagged molecule in the brain was achieved three days after injecting Incubation Day 14 eggs, suggesting that it bioaccumulates in vivo. This new immunoassay technique used to detect the novel compound in tissue samples demonstrated good repeatability.